Methods and compositions for promoting the neurological development of an infant

ABSTRACT

The present invention relates to methods for promoting the neurological development of an infant by administration of fatty acids and compositions comprising same, wherein the fatty acids are enriched with respect to docosahexaenoic acid (DHA) content.

This application is a continuation of U.S. patent application Ser. No.12/761,730, filed on Apr. 16, 2010, pending, which claims the benefit ofU.S. Provisional Application No. 61/170,685, filed on Apr. 20, 2009.Each of the above referenced applications is incorporated by referenceherein in its entirety.

TECHNICAL FIELD

The present invention is directed to methods for promoting theneurological development of an infant by administration of fatty acidsand compositions comprising same, wherein the fatty acids are enrichedwith respect to docosahexaenoic acid content.

BACKGROUND OF THE INVENTION

Preterm infants and in particular infants born prior to 33 weeksgestation are at an increased risk for developmental disorders andlearning disabilities. Long-term outcome studies of preterm infants showan overall reduction in developmental quotient (DQ) and a poorerperformance on tests of visual-motor integration, spatial relations,quantitative concepts and classroom behaviour compared with referencenorms. Such studies suggest that preterm infants may face significanteducational and developmental challenges during adolescence and also inlater life. There is therefore a need for methods by which the risk ofdevelopmental disorders and learning disabilities in preterm infants canbe minimised or even eliminated.

An inadequate nutrient supply in the neonatal period is hypothesized tocontribute to the observed poor developmental outcome in preterminfants. The n-3 long chain polyunsaturated fatty acid, docosahexaenoicacid (DHA) is of particular interest in this regard because it is amajor lipid in the brain with specific structural and functional rolesin neurological development. The uptake of DHA into the developing brainis maximised during the final trimester of pregnancy and as a resultpreterm infants do not receive the DHA in utero that is received bytheir full term counterparts.

DHA is known to significantly alter a number of basic properties of cellmembranes including permeability, fluidity and interactions withregulatory proteins. One such property includes a modulating effect onthe activity of ion channels which may facilitate electrical signalling,cellular communication and possibly brain functions such as memory,processing and the ability to learn.

The present inventors have surprisingly discovered that the neurologicaldevelopment of an infant can be promoted by administration of anincreased daily amount of DHA compared to that considered previously.

SUMMARY OF THE INVENTION

In a first aspect, the present invention provides a method for promotingthe neurological development of an infant, the method comprisingadministration to the infant of DHA in an amount of at least about 30mg/kg of body weight per day.

The DHA may be administered in an amount of at least about 60 mg/kg ofbody weight per day.

The DHA may be administered in an amount of at least about 90 mg/kg ofbody weight per day.

The infant may be a preterm infant.

The infant may be born prior to 33 weeks gestation, or prior to 36 weeksgestation.

The administration may be continued until the infant reaches termcorrected age.

The infant may be classified as small for gestational age.

The infant may have a birth weight of less than or equal to about 1250g.

The administration of DHA may commence within about 24 hours of thebirth of the infant.

The administration of DHA may commence at a time when the infantcommences enteral feeding.

The administration may be enteral or parenteral administration.

The DHA may be administered to the infant in combination with a sourceof protein.

The DHA may be administered to the infant in combination with vitaminsand/or minerals.

The DHA may be administered to the infant in combination with one ormore of the following: human milk, infant formula and human milkfortifier.

The DHA may be administered in the form of an emulsion.

The infant may be a female.

The infant may be in utero.

The administration of DHA may commence when the infant is in utero andcontinue after birth of the infant.

The infant's mother may have been identified as being at risk of givingbirth to the infant preterm, or at risk of giving birth to an infantthat will be classified as small for gestational age.

In a second aspect, the present invention provides a method forpromoting the neurological development of an infant, the methodcomprising administration to the infant fatty acids including DHA,wherein the DHA represents about 1% or more of the fatty acids.

The DHA may be present in an amount of more than 1% of the fatty acids.

The DHA may present in an amount between more than 1% and about 30% ofthe fatty acids.

The infant may be a preterm infant.

The infant may be born prior to 33 weeks gestation, or prior to 36 weeksgestation.

The administration of DHA may be continued until the infant reaches termcorrected age.

The infant may be classified as small for gestational age.

The infant may have a birth weight of less than or equal to about 1250g.

The administration of DHA may commence within about 24 hours of thebirth of the infant.

The administration of DHA may commence at a time when the infantcommences enteral feeding.

The administration may be enteral or parenteral administration.

DHA may be administered to the infant at least once a day.

DHA may be administered to the infant at least three times a day.

DHA may be administered to the infant at least five times a day.

The DHA may be administered to the infant in combination with a sourceof protein.

The DHA may be administered to the infant in combination with vitaminsand/or minerals.

The DHA may be administered to the infant in combination with one ormore of the following: human milk, infant formula and human milkfortifier.

The DHA may be administered in the form of an emulsion.

The infant may be a female.

The infant may be in utero.

The administration of DHA may commence when the infant is in utero andcontinues after birth of the infant.

The infant's mother may been identified as being at risk of giving birthto the infant preterm, or at risk of giving birth to an infant that willbe classified as small for gestational age.

In a third aspect, the present invention provides a method of feeding aninfant, the method comprising:

-   -   (i) administering fatty acids including DHA to the infant's        mother in an amount sufficient to provide a content of DHA in        the breast milk of the infant's mother that is about 1% or more        of the total fatty acids present in the breast milk, and    -   (ii) feeding the infant with breast milk following step (i).

Step (i) may comprise administering fatty acids including DHA to theinfant's mother in an amount and over a time period sufficient toprovide a constant DHA content in the breast milk of the infant's motherthat is about 1% or more of the total fatty acids present in the breastmilk of the infant's mother.

Step (i) may comprise administering fatty acids including DHA to theinfant's mother on a daily basis, and wherein the daily dosage of DHA isgreater than about 600 mg.

Step (i) may comprise administering fatty acids including DHA to theinfant's mother on a daily basis, and wherein the daily dosage of DHA isgreater than about 800 mg.

Step (i) may comprise administering fatty acids including DHA to theinfant's mother on a daily basis, and wherein the daily dosage of DHA isabout 900 mg or greater.

The infant may be a pre-term infant.

The infant may be born prior to 33 weeks gestation, or prior to 36 weeksgestation.

The feeding may be continued until the infant reaches term correctedage.

The infant may be classified as small for gestational age.

The infant may have a birth weight of less than or equal to about 1250g.

The feeding may commence within about 24 hours of the birth of theinfant.

Step (ii) may be commenced about 1 week after commencement of step (i).

The feeding may be carried out at least once a day.

The feeding may be carried out at least three times a day.

The feeding may be carried out at least five times a day.

The infant may be a female.

In a fourth aspect, the present invention provides an infant formulacomprising fatty acids including DHA, wherein the DHA is present in anamount of about 1% or more of the total fatty acid content in theformula.

The formula may be specialised preterm infant formula.

In a fifth aspect, the present invention provides a compositioncomprising fatty acids including DHA, wherein the DHA is present in anamount of about 1% or more of the total fatty acid content in thecomposition.

The composition may be a unit dosage form.

The composition may be adapted for parenteral administration.

The composition may further comprise one or more carriers, diluentsand/or adjuvants.

DEFINITIONS

In the context of the present specification, the terms “a” and “an” areused herein to refer to one or to more than one (i.e. to at least one)of the grammatical object of the article. By way of example, “anelement” means one element or more than one element.

In the context of the present specification, the term “comprising” means“including principally but not necessarily solely”. Furthermore,variations of the word “comprising”, such as “comprise” and “comprises”,have correspondingly varied meanings.

In the context of this specification, the term “about” is understood torefer to a range of numbers that a person of skill in the art wouldconsider equivalent to the recited value in the context of achieving thesame function or result.

In the context of this specification reference to “x % of the totalfatty acid content of the composition” or “x % of the total fatty acidcontent of the formula” or “x % of the total fatty acid content of themixture” and the like means x % by weight of the total fatty acidcontent of the composition, formula or mixture.

In the context of this specification, the term “neurologicaldevelopment” generally refers to growth of the brain and neurologicalsystem and to the ability of the infant to assimilate and processinformation. Neurological development may be assessed by any suitablemeans and/or criteria. Assessment may be biochemical, psychologicaland/or behavioural. One suitable means of assessing neurologicaldevelopment is the Bayley Mental Development Index (MDI).

In the context of this specification, the term “infant” means a beingthat is less than 24 months of age, and is not limited to a human. Theterm “infant” is therefore to be construed as encompassing animals, andin particular mammals, including placental mammals, monotremes andmarsupials.

In the context of this specification, the term “preterm infant” as itrelates to a human being means an infant that is born prior to 37 weeksgestation.

In the context of this specification, the term “preterm infant” as itrelates to a non-human being means an infant that is born after theperiod of viability, but before full term.

In the context of this specification, the term “small for gestationalage” means an infant whose birth weight lies below the tenth percentilefor that gestational age.

In the context of the present specification, the term “term correctedage” as it relates to a preterm human being means an age correspondingto between 37 and 40 weeks gestation had the being achieved full term.For example, if a human being is born at 30 weeks gestation, then the“term corrected age” will be reached between 7 and 10 weeks after birth.

In the context of the present specification, the term “specialisedpreterm infant formula” means an infant formula intended foradministration to preterm infants only comprising selected ingredientsso as to satisfy the unique nutritional requirements of preterm infants.

In the context of this specification, the terms “infant formula” and“infant formulas” include formulas that are intended as breast milkreplacements or supplements and also milk fortifiers.

DETAILED DESCRIPTION OF THE INVENTION

As exemplified herein, the administration of DHA and fatty acids asdescribed in accordance with the present invention has been shown tohave a positive effect on a number of indicators of neurologicaldevelopment as assessed by the Bayley Mental Development Index.Accordingly, particular embodiments of the invention relate to methodsof promoting neurological development. In addition, as also exemplifiedherein the administration of DHA and fatty acids as described is shownto reduce a number of clinical manifestations of allergy, includinghayfever. Thus, methods of the invention also find application in thetreatment or prevention of allergic conditions such as hayfever.

In a first aspect, the present invention relates to a method forpromoting the neurological development of an infant, the methodcomprising administration to the infant of DHA in an amount of at leastabout 30 mg/kg of body weight per day.

In a second aspect, the present invention relates to a method forpromoting the neurological development of an infant, the methodcomprising administration to the infant fatty acids including DHA,wherein the DHA represents about 1% or more of the fatty is acids.

The methods of the invention promote neurological development ininfants. A promotion of neurological development may be manifest byimprovements in one or more measures of neurological development such asmemory, learning and language and/or reductions in developmental delaysand learning disabilities. Those skilled in the art will also appreciatethat promoting neurological development may comprise preventing ortreating neurological or developmental disorders or conditions, forexample seizures and mental retardation.

In one embodiment, the method of the first aspect may compriseadministration to the infant of DHA in an amount of at least 30 mg/kg ofbody weight per day, at least 32 mg/kg of body weight per day, at least34 mg/kg of body weight per day, at least 36 mg/kg of body weight perday, at least 38 mg/kg of body weight per day, at least 40 mg/kg of bodyweight per day, at least 42 mg/kg of body weight per day, at least 44mg/kg of body weight per day, at least 46 mg/kg of body weight per day,at least 48 mg/kg of body weight per day, at least 50 mg/kg of bodyweight per day, at least 52 mg/kg of body weight per day, at least 54mg/kg of body weight per day, at least 56 mg/kg of body weight per day,at least 58 mg/kg of body weight per day, at least 60 mg/kg of bodyweight per day, at least 62 mg/kg of body weight per day, at least 64mg/kg of body weight per day, at least 66 mg/kg of body weight per day,at least 68 mg/kg of body weight per day, at least 70 mg/kg of bodyweight per day, at least 72 mg/kg of body weight per day, at least 74mg/kg of body weight per day, at least 76 mg/kg of body weight per day,at least 78 mg/kg of body weight per day, at least 80 mg/kg of bodyweight per day, at least 82 mg/kg of body weight per day, at least 84mg/kg of body weight per day, at least 86 mg/kg of body weight per day,at least 88 mg/kg of body weight per day, at least 90 mg/kg of bodyweight per day, at least 92 mg/kg of body weight per day, at least 94mg/kg of body weight per day, at least 96 mg/kg of body weight per day,at least 98 mg/kg of body weight per day, at least 100 mg/kg of bodyweight per day, at least 102 mg/kg of body weight per day, at least 104mg/kg of body weight per day, at least 106 mg/kg of body weight per day,at least 108 mg/kg of body weight per day, or at least 110 mg/kg of bodyweight per day.

In another embodiment, the method of the first aspect may compriseadministration to the infant of DHA in an amount between about 30 mg/kgof body weight per day and about 500 mg/kg of body weight per day, orbetween about 32 mg/kg of body weight per is day and about 500 mg/kg ofbody weight per day, or between about 34 mg/kg of body weight per dayand about 500 mg/kg of body weight per day, or between about 36 mg/kg ofbody weight per day and about 500 mg/kg of body weight per day, orbetween about 38 mg/kg of body weight per day and about 500 mg/kg ofbody weight per day, or between about 40 mg/kg of body weight per dayand about 500 mg/kg of body weight per day, or between about 42 mg/kg ofbody weight per day and about 500 mg/kg of body weight per day, orbetween about 44 mg/kg of body weight per day and about 500 mg/kg ofbody weight per day, or between about 46 mg/kg of body weight per dayand about 500 mg/kg of body weight per day, or between about 48 mg/kg ofbody weight per day and about 500 mg/kg of body weight per day, orbetween about 50 mg/kg of body weight per day and about 500 mg/kg ofbody weight per day, or between about 52 mg/kg of body weight per dayand about 500 mg/kg of body weight per day, or between about 54 mg/kg ofbody weight per day and about 500 mg/kg of body weight per day, orbetween about 56 mg/kg of body weight per day and about 500 mg/kg ofbody weight per day, or between about 58 mg/kg of body weight per dayand about 500 mg/kg of body weight per day, or between about 60 mg/kg ofbody weight per day and about 500 mg/kg of body weight per day, orbetween about 62 mg/kg of body weight per day and about 500 mg/kg ofbody weight per day, or between about 64 mg/kg of body weight per dayand about 500 mg/kg of body weight per day, or between about 66 mg/kg ofbody weight per day and about 500 mg/kg of body weight per day, orbetween about 68 mg/kg of body weight per day and about 500 mg/kg ofbody weight per day, or between about 70 mg/kg of body weight per dayand about 500 mg/kg of body weight per day, or between about 72 mg/kg ofbody weight per day and about 500 mg/kg of body weight per day, orbetween about 74 mg/kg of body weight per day and about 500 mg/kg ofbody weight per day, or between about 76 mg/kg of body weight per dayand about 500 mg/kg of body weight per day, or between about 78 mg/kg ofbody weight per day and about 500 mg/kg of body weight per day, orbetween about 80 mg/kg of body weight per day and about 500 mg/kg ofbody weight per day, or between about 82 mg/kg of body weight per dayand about 500 mg/kg of body weight per day, or between about 84 mg/kg ofbody weight per day and about 500 mg/kg of body weight per day, orbetween about 86 mg/kg of body weight per day and about 500 mg/kg ofbody weight per day, or between about 88 mg/kg of body weight per dayand about 500 mg/kg of body weight per day, or between about 90 mg/kg ofbody weight per day and is about 500 mg/kg of body weight per day, orbetween about 92 mg/kg of body weight per day and about 500 mg/kg ofbody weight per day, or between about 94 mg/kg of body weight per dayand about 500 mg/kg of body weight per day, or between about 96 mg/kg ofbody weight per day and about 500 mg/kg of body weight per day, orbetween about 98 mg/kg of body weight per day and about 500 mg/kg ofbody weight per day, or between about 100 mg/kg of body weight per dayand about 500 mg/kg of body weight per day, or between about 30 mg/kg ofbody weight per day and about 300 mg/kg of body weight per day, orbetween about 35 mg/kg of body weight per day and about 300 mg/kg ofbody weight per day, or between about 40 mg/kg of body weight per dayand about 300 mg/kg of body weight per day, or between about 45 mg/kg ofbody weight per day and about 300 mg/kg of body weight per day, orbetween about 50 mg/kg of body weight per day and about 300 mg/kg ofbody weight per day, or between about 55 mg/kg of body weight per dayand about 300 mg/kg of body weight per day, or between about 60 mg/kg ofbody weight per day and about 300 mg/kg of body weight per day, orbetween about 65 mg/kg of body weight per day and about 300 mg/kg ofbody weight per day, or between about 70 mg/kg of body weight per dayand about 300 mg/kg of body weight per day, or between about 75 mg/kg ofbody weight per day and about 300 mg/kg of body weight per day, orbetween about 80 mg/kg of body weight per day and about 300 mg/kg ofbody weight per day, or between about 85 mg/kg of body weight per dayand about 300 mg/kg of body weight per day, or between about 90 mg/kg ofbody weight per day and about 300 mg/kg of body weight per day, orbetween about 95 mg/kg of body weight per day and about 300 mg/kg ofbody weight per day, or between about 100 mg/kg of body weight per dayand about 300 mg/kg of body weight per day, or between about 30 mg/kg ofbody weight per day and about 200 mg/kg of body weight per day, orbetween about 40 mg/kg of body weight per day and about 200 mg/kg ofbody weight per day, or between about 50 mg/kg of body weight per dayand about 200 mg/kg of body weight per day, or between about 60 mg/kg ofbody weight per day and about 200 mg/kg of body weight per day, orbetween about 70 mg/kg of body weight per day and about 200 mg/kg ofbody weight per day, or between about 80 mg/kg of body weight per dayand about 200 mg/kg of body weight per day, or between about 90 mg/kg ofbody weight per day and about 200 mg/kg of body weight per day, betweenabout 60 mg/kg of body weight per day and about 180 mg/kg of body weightper day, between about 60 mg/kg of body weight per day and about 150 ismg/kg of body weight per day, or between about 60 mg/kg of body weightper day and about 120 mg/kg of body weight per day.

In another embodiment, the method of the first aspect may compriseadministration to the infant of DHA in an amount of about 30, 40, 50,60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200,210, 220, 230, 240, 250, 260, 270, 280, 290 or 300 mg/kg of body weightper day.

In the method of the second aspect, the DHA may represent about 1.05% ormore, about 1.1% or more, about 1.2% or more, about 1.3% or more, about1.4% or more, about 1.5% or more, about 1.6% or more, about 1.7% ormore, about 1.8% or more, about 1.9% or more, about 2.0% or more, about2.1% or more, about 2.2% or more, about 2.3% or more, about 2.4% ormore, about 2.5% or more, about 2.6% or more, about 2.7% or more, about2.8% or more, about 2.9% or more, about 3.0% or more, about 3.1% ormore, about 3.2% or more, about 3.3% or more, about 3.4% or more, about3.5% or more, about 3.6% or more, about 3.7% or more, about 3.8% ormore, about 3.9% or more, about 4.0% or more, about 4.1% or more, about4.2% or more, about 4.3% or more, about 4.4% or more, about 4.5% ormore, about 4.6% or more, about 4.7% or more, about 4.8% or more, about4.9% or more, or about 5.0% or more of the fatty acids.

In the method of the second aspect, the DHA may represent between morethan 1% and about 80%, or between more than 1% and about 70%, or betweenmore than 1% and about 60%, or between more than 1% and about 50%, orbetween more than 1% and about 40%, or between more than 1% and about30%, or between more than 1% and about 20%, or between more than 1% andabout 15%, or between more than 1% and about 10%, or between more than1% and about 8%, or between more than 1% and about 6%, or between morethan 1% and about 5%, or between more than 1% and about 4%, or betweenmore than 1% and about 3%, or between more than 1% and about 2%, of thefatty acids.

The infant may be a preterm infant. In an alternative embodiment theinfant may be an infant classified as small for gestational age. Infantsthat are classified as small for gestational age are typically growthrestricted in utero and hence may not have received an adequate supplyof DHA. Such infants may therefore derive significant benefit from themethods of the present invention.

The preterm infant may be born prior to 37 weeks gestation, or prior to36 weeks gestation, or prior to 35 weeks gestation, or prior to 34 weeksgestation, or prior to 33 is weeks gestation, or prior to 32 weeksgestation, or prior to 31 weeks gestation, or prior to 30 weeksgestation, or prior to 29 weeks gestation, or prior to 28 weeksgestation, or prior to 27 weeks gestation, or prior to 26 weeksgestation, or prior to 25 weeks gestation, or prior to 24 weeksgestation, or prior to 23 weeks gestation.

The methods and compositions described herein find particularapplication in relation to infant mammals, for example infant humans.However those skilled in the art will appreciate that the scope of theinvention is not so limited and that the compositions and formulasdescribed herein may be administered to infants of any species for thepurposes of nutrition and promoting neurological development, such as inlaboratory animals, domestic pets, livestock, the young of stud animalsand in rare or endangered species, for example as part of conservationmeasures in zoological parks and gardens.

The DHA may be administered to the infant via an enteral route or aparenteral route depending on the chronological age of the infant,general health of the infant and whether the infant has commencedenteral feeding. Where the infant has not yet commenced enteral feedingthe DHA may be administered parenterally, for example intravenously.Where the infant has commenced enteral feeding the DHA may beadministered enterally, for example orally. The method of enteraladministration selected will often depend on chronological age, birthweight and the clinical condition of the infant. Typically the route ofenteral administration is determined by the infant's ability toco-ordinate sucking, swallowing and breathing. Infants who are lessmature, weak or suffering from an illness may require enteraladministration via tube or possibly nasogastrically or orogastrically.

Administration of DHA may commence as soon as possible after the birthof the infant. For example, administration may commence within about 24hours of the birth of the infant, or within about 12 hours of the birthof the infant, or within about 10 hours of the birth of the infant, orwithin about 8 hours of the birth of the infant, or within about 6 hoursof the birth of the infant, or within about 5 hours of the birth of theinfant, or within about 4 hours of the birth of the infant, or withinabout 3 hours of the birth of the infant, or within about 2 hours of thebirth of the infant, or within about 1 hour of the birth of the infant.

In another embodiment, administration to the infant may commence at atime when the infant commences enteral feeding. For example, DHA may beadministered within about 24 hours, or within about 12 hours, or withinabout 8 hours, or within about 4 hours, is or within about 2 hours ofthe infant commencing enteral feeding. In an alternative embodimentadministration of DHA may commence simultaneously when the infantcommences enteral feeding.

In a further embodiment, administration may begin while the infant isstill in utero. For example, the infant's mother may be identified asbeing at risk of delivering the infant preterm (risk factors may includesmoking), or the infant's mother may have been scheduled to give birthby caesarean section. Alternatively or in addition, the infant whilst inutero may be identified as being likely to be classified as small forgestational age at birth. In such circumstances it may be desirable tobegin administration of fatty acids in accordance with methods of theinvention prior to birth of the infant. Such administration will thentypically continue beyond birth, as described herein.

The commencement of DHA administration may be determined on acase-by-case basis and could depend on a number of factors including forexample the number of weeks gestation and the general health of theinfant post birth.

Where the infant is a preterm infant administration may continue untilthe infant reaches term corrected age. For example, if the infant isborn at 32 weeks gestation administration of DHA may be continued untilthe infant reaches what would have been full term, i.e. between 37 and40 weeks gestation, or in other words until the infant reaches achronological age of between 5 and 8 weeks. Alternatively, if desired,termination of administration may occur at anytime prior to the infantreaching term corrected age.

In alternative embodiments DHA may be administered prior to and beyondterm corrected age, for example prior to, and up to 24 months after,term corrected age; or prior to, and up to 18 months after, termcorrected age; or prior to, and up to 12 months after, term correctedage; or prior to, and up to 6 months after, term corrected age.

Where the infant is a full term infant and classified as small forgestational age, administration may also commence at a time when theinfant commences enteral feeding and may continue for up to 24 monthsafter birth, or up to 18 months after birth, or up to 12 months afterbirth, or up to 9 months after birth, or up to 6 months after birth, orup to 3 months after birth.

Fatty acids administered in accordance with methods described herein maybe administered as part of a composition. Such compositions may take anumber of forms that are well known to those skilled in the artincluding tablets, capsules, caplets, powders, is solutions, suspensionsand emulsions. The form of the composition is not critical to theinvention as long as the infant receives the required amount of DHA. Theform of the composition will depend on the intended route ofadministration. Those skilled in the art will readily appreciate that anumber of suitable processes and techniques exist for the manufacture ofcompositions suitable for enteral and parenteral administration and thatthe invention is not limited by reference to any one particular processor technique. The methods may involve administration of the compositionto the infant once, twice, three, four, five, six or more times per day.

The compositions may be administered to the infant in combination with asource of protein. Protein sources are well known to those skilled inthe art and include milk, whey protein, casein, vegetable protein,animal protein, cereal protein, hydrolysed protein, amino acids,peptides and the like. In the case of humans, the source of protein maybe human milk, infant formula, human milk fortifier or combinationsthereof. The composition may also be administered to an infant incombination with various vitamins and/or minerals which are commonlygiven to preterm infants and infants classified as small for gestationalage. Examples of such vitamins and minerals include, but are not limitedto: vitamin A, B group vitamins (for example vitamins B₁, B₂, B₅, B₆, B₉and B₁₂), vitamin C, vitamin D, vitamin E, vitamin K, vitamin H, zinc,selenium, calcium, phosphorus, sodium, potassium, chloride, manganese,phosphorus, iodine, copper, iron, magnesium, molybdenum and chromium.

In the methods of the invention the DHA may be administered as part ofan infant formula. The compositions used in the method of the secondaspect may be commercially available infant formulas (for exampleformulas intended for administration to preterm infants) that have beensupplemented such that DHA is present in an amount of about 1% or moreof the total fatty acid content of the formula (presently availableinfant formulas typically comprise DHA in an amount of about 0.3% of thetotal fatty acid content). The DHA supplementation may occur duringmanufacture of the formula or alternatively post manufacture. DHAsupplementation during manufacture may be performed by replacing theusual fatty acid source or sources with an alternative fatty acid sourcehaving the desired DHA content as a percentage of total fatty acids.Alternatively, purified DHA may simply be added to the formula eitherduring or after manufacture so as to achieve the desired DHA content asa percentage of total fatty acids.

The method of the second aspect may involve administering theDHA-supplemented formula according to the daily dosage regime specifiedby the manufacturer of the formula so that the infant convenientlyreceives the specified amount of DHA as a percentage of total fatty acidcontent at each feed. Suitable infant formulas that may be supplementedand administered in this manner include but are not limited to: S-26 LBWGold available from Wyeth Nutrition, Baulkham Hills, NSW Australia,Nutriprem available from Nutricia, Macquarie Park, NSW Australia,preterm formulas sold under the trade name Enfamil® by MeadJohnson,Indiana, USA, and preNAN® available from Nestle Australia Ltd, Rhodes,NSW Australia.

In an alternative embodiment, the method of the second aspect mayinvolve the use of a DHA-supplemented commercial human milk fortifierwhich may be intended for administration to preterm infants and whichcomprises an amount of DHA such that when the fortifier is admixed withan appropriate amount of breast milk, the amount of DHA in the resultingmixture is about 1% or more of the total fatty acid content of themixture. Typically the amount of DHA added to the human milk fortifierwould be calculated on the assumption that mothers consume a minimalamount of DHA-containing products so as to ensure that when the humanmilk fortifier is added to different breast milks having a range ofdifferent DHA contents, the amount of DHA in the resultant mixtures willalways be about 1% or more of the total fatty acid content of themixture. For example, the average amount of DHA as a percentage of totalfatty acids in the milk of mothers consuming a typical western diet isless than 0.3%. Accordingly, the amount of DHA present in theDHA-supplemented commercial human milk fortifier would be an amountsufficient to increase this percentage to about 1% or more when admixedwith an appropriate amount of breast milk. Suitable commercial humanmilk fortifiers that may be supplemented and administered in the mannerdescribed include but are not limited to: products sold under the tradename Similac® by Abbott Nutrition, Illinois USA, fortifiers sold underthe trade name Enfamil® by MeadJohnson, Indiana, USA, S-26 HMF availablefrom Wyeth Nutrition, Baulkham Hills, NSW Australia, and FM-85 availablefrom Nestle Australia Ltd, Rhodes, NSW Australia.

The method of the first aspect may involve administration to the infantof purified DHA, DHA that is present as part of a mixture of fatty acidsor any composition comprising DHA (for example infant formulas and humanmilk fortifiers), so long as the is daily dosage of DHA received by theinfant is within the dosage regimes described herein.

The source of DHA in the methods, compositions and formulas describedherein may be any source known in the art, including plant and marinesources. The plant and marine sources may be genetically modified ornon-genetically modified. Marine sources include, but are not limitedto: crustaceans such as krill, molluscs such as oysters, and fish suchas salmon, trout, sardines, tuna, mackerel, sea bass, menhaden, herring,pilchards, kipper, eel or whitebait. The DHA may be present in apurified form and/or in the form of an extract from a suitable source.The DHA may be present as a component of fish oil. The fish oil may beobtained from, for example one or more of the following fish: tuna,salmon, trout, menhaden, sea bass, mackerel, sardines, pilchards,herring, kipper, eel, whitebait or any other “fatty fish”.

The DHA may be stabilised so as to protect against oxidation and otherforms of degradation. The stabilisation may be achieved byencapsulation. Suitable encapsulation methods include, but are notlimited to: coatings (including primary, secondary and tertiary),emulsions, coacervasion and gels. The DHA may alternatively bestabilised in the form of an emulsion wherein the DHA is either not, oronly partially encapsulated.

Preterm human infants and human infants classified as small forgestational age are typically able to consume milk and/or formula in anamount between about 100 mL and 250 mL per kg of body weight per day.Accordingly, the method of the second aspect may comprise administrationto the infant of milk and/or formula in an amount between about 100 mLand 250 mL per kg of body weight per day, wherein the DHA content of themilk or formula is about 1% or more of the total fatty acid content ofthe milk or formula. The milk and/or formula may be administered to theinfant once a day or on multiple occasions depending on the infant'sage, general health and feeding regime. As the infant grows, the amountof milk and/or formula can be increased in accordance with the infant'snutritional requirements.

In a third aspect, the present invention relates to a method of feedingan infant, the method comprising:

-   -   (i) administering fatty acids including DHA to the infant's        mother in an amount sufficient to provide a content of DHA in        the breast milk of the infant's mother that is about 1% or more        of the total fatty acids present in the breast milk, and    -   (ii) feeding the infant with breast milk following step (i).

The infant may be a preterm infant or an infant classified as small forgestational age. The feeding may comprise breast feeding oralternatively bottle feeding using expressed milk from the breast of theinfant's mother.

Step (i) may comprise administering fatty acids including DHA to theinfant's mother in an amount sufficient to provide a content of DHA inthe breast milk of the infant's mother that is at least 1%, or at least1.05%, or at least 1.1%, or at least 1.2%, or at least 1.3%, or at least1.4%, or at least 1.5%, or at least 1.6%, or at least 1.7%, or at least1.8%, or at least 1.9%, or at least 2%, or at least 2.1%, or at least2.2%, or at least 2.3%, or at least 2.4%, or at least 2.5%, or at least2.6%, or at least 2.7%, or at least 2.8%, or at least 2.9%, or at least3.0% of the total fatty acids present in the breast milk.

Step (i) may comprise administering fatty acids including DHA to theinfant's mother in an amount sufficient to provide a content of DHA inthe breast milk of the infant's mother that is between more than 1% andabout 20%, or between more than 1.05% and about 20%, or between morethan 1.1% and about 20%, or between more than 1% and about 30%, orbetween more than 1% and about 25%, or between more than 1% and about15%, or between more than 1% and about 10%, or between about 1.5% andabout 20%, or between about 1.5% and about 15%, or between about 1.5%and about 10%, or between about 1.5% and about 5%, or between about 2%and about 20%, or between about 2% and about 15%, or between about 2%and about 10%, of the total fatty acids present in the breast milk.

Step (i) may comprise administering fatty acids including DHA to theinfant's mother in an amount and over a time period sufficient toprovide a constant DHA content in the breast milk of the infant's motherthat is about 1% or more of the total fatty acids present in the breastmilk of the infant's mother. This outcome may be achieved byadministering to the infant's mother approximately 600 mg or more of DHAper day. However, in the case of a mother who consumes minimal DHA aspart of her diet, the amount of DHA administered on a daily basis may beapproximately 900 mg or more. In the case of a mother who consumes adiet rich in DHA, it may be possible to achieve a content of DHA in thebreast milk of the infant's mother that is about 1% or more of the totalfatty acids present by administering less than 600 mg of DHA per day.Those skilled in the art will be capable of determining the amount ofDHA required to be administered to is an infant's mother in order toachieve a desired amount of DHA as a percentage of total fatty acids inbreast milk by routine trial and experimentation based on the teachingsherein.

In one embodiment, the amount of DHA administered to the infant's mothermay be between about 600 mg per day and about 4000 mg per day, orbetween about 600 mg per day and about 3500 mg per day, or between about600 mg per day and about 3000 mg per day, or between about 600 mg perday and about 2500 mg per day, or between about 600 mg per day and about2000 mg per day, or between about 600 mg per day and about 1500 mg perday, or between about 600 mg per day and about 1200 mg per day.

The DHA may be administered in multiple unit dosage forms (such astablets or capsules for example), or alternatively in a single unitdosage form comprising the daily amount of DHA. The DHA may beadministered in the form of DHA-rich tuna oil, for example the productsold under the trade name HiDHA® available from Nu-Mega Ingredients,Nathan, Queensland Australia. In manipulating the amount of DHA presentin the breast milk, the naturally occurring amount of arachadonic acidpresent (about 0.4% to 0.6%) may not be substantially altered.

Step (ii) may be commenced at any time after step (i). In one embodimentstep (ii) commences about 1 week after commencement of step (i) therebyensuring that the desired amount of DHA in the breast milk has beenreached and is essentially constant. Practically however it is likelythat the infant's mother will simply commence step (i) whilst continuingthe infant's standard feeding regime. Accordingly, the level of DHA inthe breast milk being received by the infant will increase andsubsequently be maintained at, or above, the desired level.

The method of the third aspect may commence at anytime after the birthof the infant, and in one embodiment within about 24 hours of the birthof the infant. Commencement of the method will of course be dependent onwhether the infant is capable of breastfeeding or bottle feedingexpressed milk. Where the infant is unable to commence breastfeeding orbottle feeding soon after birth, the method may be commenced once theinfant is able to breastfeed or bottle feed successfully.

In a fourth aspect, the present invention relates to infant formulascomprising fatty acids including DHA, wherein the DHA is present in anamount of about 1% or more of the total fatty acid content in theformulas. The infant formulas may be milk fortifiers or is formulasintended for use as breast milk replacements or supplements.

The DHA may be present in the formulas in an amount of about 1.05% ormore, about 1.1% or more, about 1.2% or more, about 1.3% or more, about1.4% or more, about 1.5% or more, about 1.6% or more, about 1.7% ormore, about 1.8% or more, about 1.9% or more, about 2.0% or more, about2.1% or more, about 2.2% or more, about 2.3% or more, about 2.4% ormore, about 2.5% or more, about 2.6% or more, about 2.7% or more, about2.8% or more, about 2.9% or more, about 3.0% or more, about 3.1% ormore, about 3.2% or more, about 3.3% or more, about 3.4% or more, about3.5% or more, about 3.6% or more, about 3.7% or more, about 3.8% ormore, about 3.9% or more, about 4.0% or more, about 4.1% or more, about4.2% or more, about 4.3% or more, about 4.4% or more, about 4.5% ormore, about 4.6% or more, about 4.7% or more, about 4.8% or more, about4.9% or more, or about 5.0% or more, of the total fatty acid content ofthe formulas.

In alternative embodiments, the DHA may be present in the formulas in anamount between more than 1% and about 80%, or between more than 1% andabout 70%, or between more than 1% and about 60%, or between more than1% and about 50%, or between more than 1% and about 40%, or between morethan 1% and about 30%, or between more than 1% and about 20%, or betweenmore than 1% and about 15%, or between more than 1% and about 10%, orbetween more than 1% and about 8%, or between more than 1% and about 6%,or between more than 1% and about 5%, or between more than 1% and about4%, or between more than 1% and about 3%, or between more than 1% andabout 2%, of the total fatty acid content of the formulas.

The infant formulas in accordance with the invention will typically benutritionally complete and include standard ingredients that are presentin commercially available infant formulas and milk fortifiers, such asfat, protein, carbohydrate, vitamins and minerals. The nutritionalcomposition of the formulas and fortifiers may be adjusted depending onthe intended age group to which the formulas and fortifiers are to beadministered. For example, in one embodiment the infant formulas arespecialised preterm infant formulas and hence include selectedingredients so as to satisfy the unique nutritional requirements ofpreterm infants. Alternatively, where the formulas and fortifiers areintended to be administered to infants that are older than termcorrected age, the nutritional composition may be adjusted to meet thediffering nutritional requirements of such infants. The formulas may bein the form of liquids, powders or tablets and may be manufactured isaccording to techniques well known to those skilled in the art.

In one embodiment, the infant formulas of the invention may be preparedby supplementing commercially available infant formulas or fortifierswith the desired amount of DHA as described above in connection with thesecond aspect. Suitable infant formulas that may be supplemented in thismanner include, but are not limited to: products sold under the tradename Similac® by Abbott Nutrition, Illinois USA, products sold under thetrade name S-26 (for example S-26 LBW Gold and S-26 HMF) available fromWyeth Nutrition, Baulkham Hills, NSW Australia, Nutriprem available fromNutricia, Macquarie Park, NSW Australia, products sold under the tradename Enfamil® by MeadJohnson, Indiana, USA and FM-85 and preNAN®, bothavailable from Nestle Australia Ltd, Rhodes, NSW Australia.

In a fifth aspect, the present invention relates to a compositioncomprising fatty acids including DHA, wherein the DHA is present in anamount of about 1% or more of the total fatty acid content in thecomposition.

The DHA may be present in the composition in an amount of about 1.05% ormore, about 1.1% or more, about 1.2% or more, about 1.3% or more, about1.4% or more, about 1.5% or more, about 1.6% or more, about 1.7% ormore, about 1.8% or more, about 1.9% or more, about 2.0% or more, about2.1% or more, about 2.2% or more, about 2.3% or more, about 2.4% ormore, about 2.5% or more, about 2.6% or more, about 2.7% or more, about2.8% or more, about 2.9% or more, about 3.0% or more, about 3.1% ormore, about 3.2% or more, about 3.3% or more, about 3.4% or more, about3.5% or more, about 3.6% or more, about 3.7% or more, about 3.8% ormore, about 3.9% or more, about 4.0% or more, about 4.1% or more, about4.2% or more, about 4.3% or more, about 4.4% or more, about 4.5% ormore, about 4.6% or more, about 4.7% or more, about 4.8% or more, about4.9% or more, or about 5.0% or more, of the total fatty acid content ofthe composition.

In alternative embodiments, the DHA may be present in the composition inan amount between more than 1% and about 80%, or between more than 1%and about 70%, or between more than 1% and about 60%, or between morethan 1% and about 50%, or between more than 1% and about 40%, or betweenmore than 1% and about 30%, or between more than 1% and about 20%, orbetween more than 1% and about 15%, or between more than 1% and about10%, or between more than 1% and about 8%, or between more than 1% andabout 6%, or between more than 1% and about 5%, or between is more than1% and about 4%, or between more than 1% and about 3%, or between morethan 1% and about 2%, of the total fatty acid content of thecomposition.

The composition may comprise one or more acceptable carriers, diluentsand/or adjuvants. The carriers, diluents and adjuvants must be“acceptable” in terms of being compatible with the other components ofthe composition and not deleterious to the recipient thereof.

Suitable carriers, diluents and adjuvants will depend on the intendedroute of administration and are well known to those skilled in the art.In one embodiment, the composition may be adapted for parenteraladministration. For administration as an injectable solution orsuspension non-toxic parenterally acceptable diluents or carriers caninclude: Ringer's solution, isotonic saline, glucose solution, distilledwater and phosphate buffered saline. Methods for preparing parenterallyadministrable compositions are apparent to those skilled in the art, andare described in more detail in, for example, Remington's PharmaceuticalScience, 15th ed., Mack Publishing Company, Easton, Pa., herebyincorporated by reference herein.

The composition may be a unit dosage form suitable for oraladministration, for example a tablet, capsule, caplet, or lozenge. Unitdosage forms may optionally include one or more acceptable excipients,for example ethyl cellulose, cellulose acetate phthalates, mannitol,lactose, starch, magnesium stearate, sodium saccharin, talc, glucose,sucrose, carbonate, and the like.

The invention will now be described in more detail, by way ofillustration only, with respect to the following examples. The examplesare intended to serve to illustrate this invention and should not beconstrued as limiting the generality of the disclosure of thedescription throughout this specification.

Example Clinical Study Study Design and Methodology

A multi-centre randomized controlled trial was conducted in 5 Australianperinatal centres. Ethics approval was granted by the localInstitutional Review Boards (Human Research Ethics Committees) of eachcentre. The trial began with a pilot phase at the Women's and Children'sHospital, Adelaide. A central trial coordinator monitored datacollection, entry and checking. An independent serious adverse eventcommittee reviewed all deaths.

Infants born <33 weeks were eligible and families were approached by theresearch nurses within 5 days of their infant receiving any enteralfeeds. Infants were excluded if they had major congenital or chromosomalabnormalities, were from a multiple birth where not all live borninfants were eligible, or were in other trials of fatty acidsupplementation. Lactating mothers in whom tuna oil was contraindicated,for example bleeding disorders or therapy with anticoagulants, were alsoexcluded.

Once written informed consent was obtained, mother-infant pairs wererandomly assigned a unique study number through a computer driventelephone randomization service according to an independently generatedrandomization schedule. Stratification was by centre, birth weight(<1250 g and ≧1250 g) and infant sex. Multiple births were considered asingle randomization unit and randomization of twins or triplets wasaccording to the sex and birth weight of the first born infant. Baselinecharacteristics, including maternal age, infant race as identified byparents, parental education, birth order, parity, gestational age atbirth, birth measurements, pregnancy and birth complications, wererecorded.

Lactating mothers allocated to the high-DHA group were asked to consume6×500 mg DHA-rich tuna oil capsules per day in order to achieve a breastmilk DHA concentration that was ≈1% of total fatty acids withoutaltering the naturally occurring concentration of arachidonic acid (AA)in breast milk. If supplementary formula was required, infants weregiven a high-DHA preterm formula (≈1.0% DHA, 0.6% AA). Mothers withinfants allocated to the standard DHA group were asked to consume 6×500mg placebo, soy oil capsules that did not change the fat content orfatty acid composition of their milk. In the event that mothers chosenot to breastfeed or could not produce enough breast milk, infants werefed standard preterm infant formula (≈0.35% DHA and 0.6% AA). Tofacilitate blinding, each treatment group was separately colour codedinto two groups. All capsules were similar in size, shape and colour andwere donated by Clover Corporation, Sydney, Australia. If formula wasrequired in the pilot phase, two drops of oil from capsules in matchingcolour coded containers were added to each 90 mL jar of formula. For theremainder of the trial, Mead Johnson Nutritionals, IN, USA, specificallymanufactured ready-to-feed preterm formula to trial specifications andpackaged the formula according to the colour codes. The interventioncontinued until infants reached their expected date of delivery (EDD).Infants in the high-DHA group consumed approximately 60 mg/kg of bodyweight per day of DHA, whereas infants in the standard DHA groupconsumed approximately 20 mg/kg of body weight per day of DHA. Whilst inhospital, the feeding regimen was under the direction of the infant'sclinician and did not interfere with the use of human milk fortifier orsupplementary vitamins or minerals. Post-term, breastfeeding motherswere encouraged to continue breastfeeding and those who had weaned toformula were encouraged to use a term formula supplemented with DHA andAA. Parents were reimbursed the difference in cost betweenunsupplemented term formula and DHA-supplemented term formula.

During the intervention the proportion of parenteral and enteralnutrition, human milk and infant formula intakes, and the frequency ofinterrupted feeds were documented weekly. Confirmed cases of necrotizingenterocolitis (NEC), sepsis, intra-ventricular hemorrhage (IVH),retinopathy of prematurity (ROP) and oxygen treatment at 36 weeks werealso documented. Weight, length and head circumference were assessed atEDD and women who were breastfeeding donated a 5 mL sample of milk toassess the fatty acid composition.¹ At EDD, women were also asked toguess their group allocation.

The Mental Development Index (MDI) of the Bayley Scales of InfantDevelopment, Second Edition (BSID-II)² evaluates memory, habituation,problem solving, early number concepts and language. MDI at 18 months'corrected age was chosen as the primary outcome because it represents arobust assessment of mental delays in children, is reasonably correlatedwith IQ in preterm children³ and allows comparison with other relevantstudies. The psychomotor development index (PDI), which evaluatescontrol of the gross muscle groups including movements associated withstanding, walking, running and jumping, as well as fine motormanipulations involved in prehension, adaptive use of writingimplements, and imitation of hand movements, was a secondary outcome.MDI and PDI scores standardized to a mean of 100 with a standarddeviation of 15 (range from 50 to 150). If a child performed below thethreshold of the tests for either MDI or PDI, they were assigned a scoreof 45. If they were completely untestable because of severe delay, theywere assigned a score of 40. At the time of the BSID assessment weight,length and head circumference were measured and the Home ScreeningQuestionnaire (HSQ)⁴ was administered to assess the quality and quantityof cognitive, social and emotional support available to each infant inthe home environment. Parents, clinicians and all is research personnelwere blinded to participant study group.

The trial was designed to evaluate the effect of high dietary DHA in thepreterm period on infants born <33 weeks gestation as well as importantsubgroups in this heterogeneous population. We also planned a priori toconduct a sub-group analysis based on infant sex because DQ in earlychildhood often varies according to sex and differences of the order of5-8 points have been reported. Overall, our recruitment target was 320infants per group to allow for 10% loss to follow-up, including deaths.

All analyses were conducted according to the intention to treatprinciple. The a priori level of significance was p<0.05. Most of theoutcomes were analysed using Generalized Estimating Equations (GEE)⁵ toaccount for the clustering of infants within mother using SAS version9.1 (SAS Institute Inc., Cary, N.C., USA). Normally distributed outcomeswere analysed using a linear GEE, with the difference in means (95% CI)as the treatment effect. The subgroup analyses were performed viafactorial models to allow testing for an interaction between treatmentand subgroup. Outcomes that were counts were analysed using Poisson orNegative-Binomial GEE as appropriate, with the ratio of means (95% CI)as the treatment effect. Binary or categorical data were analysed usinglog-binomial GEE, with the relative risk (ratio of proportions) (95% CI)as the treatment effect. In secondary analyses the Bayley outcomes werealso adjusted for the potential confounders of maternal education,infant sex, gestational age at delivery and birth order. An additionaladjustment was made for phase of the study, which made little differenceto the results. All other outcomes were adjusted for the potentialconfounders of infant sex and gestational age at delivery. In post hocanalyses we also investigated whether groups differed in the proportionof children with mild (<85) and significant (<70) mental delay. Missingdata were multiply imputed using regression models (either normal,poisson or binary) with 50 imputations. Sensitivity analyses wereperformed using different seeds, increasing the number of imputations oradding further terms to the regression models. The results of thesesensitivity analyses were similar to those presented here.

Results

Enrolment for the trial began on Apr. 4, 2001 and ended Oct. 28, 2005.Follow-up commenced on Jan. 17, 2003 and ended Sep. 21, 2007. Adequatedata for the analysis of the primary outcome were available on 614infants, 93.5% of the infants who were originally enrolled in the trial(92.5% in the high-DHA group and 94.3% in the is standard-DHA group).The demographic and clinical characteristics of the infants and theirfamilies at randomization were comparable between the two groups (seeTable 1).

Median duration of treatment was comparable between the high-DHA (9.4weeks, inter-quartile range, IQR 7.9 to 11.4 weeks) and the standard-DHA(9.4 weeks, IQR 8.0 to 11.6 weeks) groups. Maternal compliance based oncapsule returns was 81.1% in the high-DHA group and 81.7% in thestandard-DHA group (p=0.88). Mean DHA (±SD) concentration in the milk ofwomen in the high-DHA group was greater than standard treatment(0.85±0.39% vs. 0.25±0.13% total fatty acids, p<0.0001) as was the meanDHA concentration in the three batches of preterm formula used for thetrial (1.11±0.29% vs. 0.42±0.05% total fatty acids, p<0.0001). The meanAA (±SD) concentration did not differ between groups for human milk(0.41±0.09% vs. 0.40±0.09% total fatty acids) or preterm infant formula(0.69±0.29% vs. 0.69±0.22% total fatty acids). At the end of dietarytreatment 72% of women in the high-DHA group correctly guessed theirgroup allocation as indicated by more frequent reports of fishyeructations from the high-DHA group compared with standard treatment(140/322 vs. 24/335, unadjusted relative risk 6.20, 95% confidenceinterval, CI, 3.79 to 10.20, p<0.0001). There were no differencesbetween the groups in maternal report of diarrhea, constipation, nauseaor vomiting.

The primary outcome of mean Bayley MDI did not differ between thehigh-DHA and standard-DHA groups (mean difference, MD, 1.9, 95% CI, −1.0to 4.7, unadjusted; MD, 1.6, 95% CI −1.2 to 4.3, adjusted). A priorisub-group analyses based on the randomization strata showed interactionsbetween dietary treatment and sex, and between dietary treatment andbirth weight. The MDI of girls fed the high-DHA diet was significantlyhigher than girls fed the standard-DHA diet in both unadjusted andadjusted analyses (MD, 4.7, 95% CI 0.5 to 8.8 unadjusted; MD, 4.5, 95%CI 0.5 to 8.5 adjusted, Table 2). The MDI of infants born <1250 g andfed a high-DHA diet to EDD was higher than that of infants fedstandard-DHA diets in the unadjusted comparison (MD, 4.7, 95% CI 0.2 to9.2). For Bayley PDI there was no significant difference between groups(MD, 0.9, 95% CI, −1.8 to 3.6). There were no interactions between dietand sex or between diet and birth weight strata and consequently nodifferences in PDI between the groups in either of the birth weightstrata or for boys and girls. At 18 months' corrected age, the degree ofsocial and cognitive stimulation available in the home environment didnot differ between is groups (HSQ score ±SD, 34±4, n=322 in the high-DHAgroup vs. 34±3, n=335 in the standard-DHA group).

Post-hoc analyses indicated that overall fewer infants had significantlydelayed mental development with high-DHA diets compared with standardDHA. There were fewer infants born <1250 g in the high-DHA group withmildly delayed mental development and fewer infants born ≧1250 g fed thehigh-DHA diet with significant mental delay compared with thestandard-DHA diet.

The secondary clinical outcomes of the infants are shown in Table 2.Blindness and hearing impairment requiring aids were rare. There were nodifferences in anthropometric measures between the groups except that at18 months' corrected age infants who were allocated the high-DHA dietwere longer than infants allocated to the standard diet (Table 2). Theextent of breastfeeding did not differ between groups at EDD (Table 2),4, 12 or 18 months corrected age. Other secondary outcomes also did notdiffer between groups, but fewer infants fed high-DHA in the pretermperiod required oxygen treatment at 36 weeks compared with standard DHAtreatment after correction for gestational age at birth and sex (Table2). There were two maternal deaths after the end of the interventionphase (suicide by hanging and Wolff Parkinson White Syndrome withsecondary substance abuse) in the standard-DHA group.

Impacts of the high-DHA and standard-DHA diet on various allergyindicators was also assessed at 12 or 18 months corrected age. As shownin Table 3, reductions in the incidence of hayfever, asthma and eczemawere all observed in infants on the high-DHA diet. The reduction inhayfever incidence was of particular significance (p-value=0.03), andwhen analysed more closely the reductions were found to be most dramaticin boys (1.6% on high-DHA compared to 10.9% on standard-DHA; p=0.01).

REFERENCES

-   1. Makrides M., Neumann M. A., Gibson R. A. Effect of maternal    docosahexaenoic acid (DHA) supplementation on breast milk    composition. Eur J Clin Nutr. 1996; 50(6): 352-357.-   2. Bayley N. Manual for the Bayley Scales of Infant Development,    Second Edition (BSID-II). San Antonio, Tex.: Psychological    Corporation; 1993.-   3. Lucas A., Morley R., Cole T. J., Lister G., Leeson-Payne C.    Breast milk and subsequent intelligence quotient in children born    preterm. Lancet. 1992; 339(8788):

261-264.

-   4. Frankenburg W. K., Coons C. E. Home Screening Questionnaire: its    validity in assessing home environment. J Pediatr. 1986; 108(4):    624-626.-   5. Liang K-Y., Zeger S. L. Longitudinal data analysis using    generalized linear models. Biometrika. 1986; 73(1): 13-22.

The reference in this specification to any prior publication (orinformation derived from it), or to any matter which is known, is not,and should not be taken as an acknowledgment or admission or any form ofsuggestion that that prior publication (or information derived from it)or known matter forms part of the common general knowledge in the fieldof endeavour to which this specification relates.

It will be appreciated by persons skilled in the art that numerousvariations and/or modifications may be made to the invention withoutdeparting from the spirit or scope of the invention as broadlydescribed. The present embodiments are therefore to be considered in allrespects as illustrative and not restrictive.

TABLE 1 Baseline Demographic and Clinical Characteristics of Childrenand their Families High-DHA Standard-DHA Characteristic (n = 322) (n =335) Recruitment Hospital Flinders Medical Centre 31 (9.6) 32 (9.6) KingEdward Memorial Hospital 65 (20.2) 57 (17.0) Royal Brisbane & Women'sHospital 46 (14.3) 50 (14.9) Royal Women's Hospital 61 (18.9) 63 (18.8)Women's & Children's Hospital 119 (37.0) 133 (39.7) Mother's Age atTrial Entry (years) 29.9 (5.8) 30.2 (5.4) Mother Completed SecondaryEducation 205 (63.7) 201 (60.1) Father Completed Secondary Education 172(53.5) 188 (56.0) Mother Smoked During Pregnancy 82 (25.6) 84 (25.1)Previous PreTerm Births 51 (15.8) 58 (17.4) Birth by Caesarean Section220 (68.3) 235 (70.0) Antenatal Steroids Administered 279 (86.6) 302(90.1) Multiple Pregnancy 98 (30.4) 123 (36.7) Gestational Age at Birth(weeks) 30 (27-31) 30 (27-31) Caucasian 283 (87.9) 311 (92.8) Male Sex173 (53.7) 182 (54.3) Birth Weight (g) 1308 (423) 1307 (415) Small forgestational age 61 (18.9) 62 (18.6) Birth Weight <1250 g 147 (45.7) 149(44.5) Recumbent Length At Birth (cm) 38.2 (4.0) 38.1 (4.0) HeadCircumference At Birth (cm) 27.2 (2.8) 27.3 (2.7) Days of PartialEnteral Feeds 2 (1-4) 2 (0-3) Pre-Randomization Infant Age atRandomization (days) 4 (3-6) 4 (2-5) Infants Receiving Breast Milk at297 (92.2) 306 (91.3) Trial Entry Values are number (%) of babies, mean(sd) or median (interquartile range)

TABLE 2 Secondary Clinical Outcomes Unadjusted Unadjusted AdjustedHigh-DHA Standard-DHA Effect (95% CI) P-value Effect (95% CI) AdjustedP-value All n = 322 n = 335 Death  9 (2.8)  9 (2.7) 1.04 (0.42, 2.59)0.93 1.09 (0.44, 2.66) 0.86 Pre-Discharge Death  9 (2.8)  6 (1.8) 1.56(0.56, 4.34) 0.39 1.66 (0.63, 4.41) 0.31 Days In NICU 22 (3-31) 21(4-33) 1.02 (0.82, 1.27) 0.87 1.03 (0.88, 1.20) 0.75 Days In HospitalCare  64 (40-80)  64 (41-80) 1.01 (0.92, 1.10) 0.87 1.00 (0.95, 1.06)0.92 Days On Parenteral Nutrition 12 (5-15) 12 (5-14) 1.06 (0.90, 1.24)0.52 1.03 (0.92, 1.16) 0.59 Days Of Intravenous Lipids  8 (0-12)  8(0-10) 1.06 (0.85, 1.32) 0.59 1.06 (0.87, 1.30) 0.54 Days Until FullEnteral Feeds 12 (6-14) 12 (6-14) −0.2 (−2.0, 1.6)  0.82 −0.2 (−1.0,0.6)  0.55 Exclusively Human Milk Fed at EDD 142 (44.1) 135 (40.2) 1.10(0.87, 1.39) 0.42 1.11 (0.88, 1.40) 0.39 Feeding Interrupted 106 (32.9)106 (31.6) 1.04 (0.82, 1.32) 0.74 1.07 (0.87, 1.31) 0.55 Any NEC 14(4.3)  7 (2.1) 2.06 (0.83, 5.13) 0.12 2.14 (0.87, 5.22) 0.10 BowelSurgery 12 (3.7)  9 (2.7) 1.39 (0.58, 3.33) 0.46 1.45 (0.63, 3.35) 0.39Oxygen Treatment at 36 weeks  60 (18.6)  84 (25.1) 0.74 (0.54, 1.02)0.07 0.76 (0.58, 1.00) 0.05 Any intraventricular Haemorrhage  45 (14.0) 44 (13.2) 1.06 (0.71, 1.59) 0.77 1.07 (0.72, 1.58) 0.73 Severeintraventricular Haemorrhage^(a)  9 (2.8)  6 (1.8) 1.56 (0.56, 4.33)0.39 1.63 (0.61, 4.33) 0.33 Any Retinopathy of Prematurity  74 (23.0) 73 (21.8) 1.05 (0.77, 1.45) 0.74 1.09 (0.85, 1.40) 0.49 SevereRetinopathy of Prematurity^(b) 14 (4.3) 17 (5.1) 0.86 (0.42, 1.75) 0.670.91 (0.46, 1.80) 0.79 Any Sepsis  53 (16.6)  48 (14.3) 1.16 (0.79,1.69) 0.46 1.18 (0.85, 1.65) 0.32 Postnatal steroids 30 (9.3)  34 (10.2)0.92 (0.56, 1.51) 0.73 0.96 (0.61, 1.50) 0.85 Small for gestational ageat EDD 109 (33.8) 105 (31.4) 1.08 (0.85, 1.37) 0.53 1.09 (0.86, 1.37)0.49 Weight at EDD (g) 3175 (553)   3129 (535)     42 (−118, 203) 0.60  42 (−116, 199) 0.60 Weight at 18 Months (g) 11625 (1811)   11277(1588)    201 (−237, 639) 0.37  187 (−250, 623) 0.40 Length at EDD (cm)48.7 (3.3)   48.4 (3.3)    0.2 (−0.3, 0.8)  0.42   0.2 (−0.3, 0.8)  0.45Length at 18 Months (cm) 82.8 (5.2)   81.7 (4.7)  0.9 (0.2, 1.7)  0.010.9 (0.2, 1.6)  0.01 Head circumference at EDD (cm) 35.4 (1.8)   35.4(1.9)    0.10 (−0.6, 0.7)  0.85   0.10 (−0.6, 0.7)  0.85 Headcircumference at 18 Months (cm) 47.6 (2.5)   47.6 (2.2)  −0.06 (−0.8,0.7)  0.86 −0.05 (−0.8, 0.7)  0.88 Seizures at 18 months  7 (2.0) 17(5.2) 0.39 (0.15, 1.04) 0.06 0.39 (0.15, 1.04) 0.06 Unilateral orbilateral blindness at 18 0  1 (0.3) — — — — months Severe hearing lossrequiring aids at 0  1 (0.3) — — — — 18 months Cerebral Palsy at 18months 13 (3.9) 10 (3.0) 1.31 (0.56, 3.06) 0.53 1.31 (0.56, 3.06) 0.53Values are number of babies (%) with effect being Relative Risk or mean(interquartile range) with Ratio of Means as effect Adjusted for GA atdelivery and sex. Further adjustment for pilot phase vs multi-centrephase did not alter the results. ^(a)Grade 3 or 4 ^(b)Grade 3 or higher

TABLE 3 Allergy diagnosis at 12 or 18 months corrected age High-DHAStandard-DHA Hayfever  3.5%  8.6% (p = 0.03) Hayfever (boys)  1.6% 10.9%(p = 0.01) Hayfever (girls)  5.7%  5.6% Asthma 19.8% 21.0% Eczema 25.8%27.0%

1. A method for promoting the neurological development of an infant, themethod comprising administration to the infant of DHA in an amount of atleast about 30 mg/kg of body weight per day.
 2. The method of claim 1,comprising administration to the infant of DHA in an amount of at leastabout 60 mg/kg of body weight per day.
 3. The method of claim 2,comprising administration to the infant of DHA in an amount of at leastabout 90 mg/kg of body weight per day.
 4. The method of claim 1, whereinthe infant is a preterm infant.
 5. The method of claim 4, wherein theinfant is born prior to 33 weeks gestation.
 6. The method of claim 4,wherein the infant is born prior to 36 weeks gestation.
 7. The method ofclaim 4, wherein the administration is continued until the infantreaches term corrected age.
 8. The method of claim 1, wherein the infantis classified as small for gestational age.
 9. The method of claim 1,wherein the infant has a birth weight of less than or equal to about1250 g.
 10. The method of claim 1, wherein the administration commenceswithin about 24 hours of the birth of the infant.
 11. The method ofclaim 1, wherein the administration commences at a time when the infantcommences enteral feeding.
 12. The method of claim 1, wherein theadministration is enteral or parenteral administration.
 13. The methodof claim 1, wherein the DHA is administered to the infant in combinationwith a source of protein.
 14. The method of claim 1, wherein the DHA isadministered to the infant in combination with vitamins and/or minerals.15. The method of claim 1, wherein the DHA is administered to the infantin combination with one or more of the following: human milk, infantformula and human milk fortifier.
 16. The method of claim 1, wherein theDHA is administered in the form of an emulsion.
 17. The method of claim1, wherein the infant is in utero.
 18. The method of claim 17, whereinthe administration commences when the infant is in utero and continuesafter birth of the infant.
 19. (canceled)
 20. A method for promoting theneurological development of an infant, the method comprisingadministration to the infant fatty acids including DHA, wherein the DHArepresents about 1% or more of the fatty acids. 21-41. (canceled)
 42. Amethod of feeding an infant, the method comprising: (i) administeringfatty acids including DHA to the infant's mother in an amount sufficientto provide a content of DHA in the breast milk of the infant's motherthat is about 1% or more of the total fatty acids present in the breastmilk, and (ii) feeding the infant with breast milk following step (i).43-64. (canceled)